Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
  • C07K14/575—Hormones
  • C07K14/585—Calcitonins
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P3/00—Drugs for disorders of the metabolism
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides

Definitions

• the subject of the invention is in particular the compounds corresponding to general formulas (I A ) and (Is), as defined above, in which As and A '7 represents a valine.
• the polypeptides according to the invention lower calcemia and phosphatemia; they inhibit bone destruction, accelerate the formation of new bones, increase the intestinal absorption of calcium and decrease calciuria. They fix the calcium inside the cells. They have a major anti-inflammatory and analgesic effect.
• the invention therefore also relates to the polypeptides of formulas (I A ) and (I B ) as defined above, as medicaments.
• a more particular subject of the invention is, as medicaments, the preferred polypeptides mentioned above.
• the main indications of the drugs according to the invention are represented by Poppy's disease, osteoporosis of various etiologies (common, poro-malacic, cortisonic, post-traumatic, immobilization and idiopathic), renal osteodystrophy, fractures, hypercalcemia, osteoarticular pain, in particular those following bone metastases, spasmophilia and normocalcemic tetany.
• the invention also relates to pharmaceutical compositions containing as active principle at least one of the medicaments as defined above.
• compositions can be, for example, solid or liquid and can be in the pharmaceutical forms commonly used in human medicine, such as, for example, simple or coated tablets, capsules, capsules, granules, liposomes, aerosols , suppositories, injections; they are prepared according to the usual methods.
• the active ingredient (s) can be incorporated therein into excipients usually used in these pharmaceutical compositions, such as talc, gum arabic, lactose, starch, magnesium stearate, cocoa butter, aqueous vehicles or not , fatty substances of animal or vegetable origin, paraffinic derivatives, glycols, various wetting agents, dispersants or emulsifiers, preservatives.
• the invention particularly relates to pharmaceutical compositions as defined above, characterized in that they are intended for parenteral, oral administration in the form of a liposome or nasal in the form of a spray.
• the subject of the invention is also a process for preparing the compounds as defined above, characterized in that amino acids, peptides or their combinations are subjected to condensation reactions in the order of the acid sequence amines of formulas (I A ) and (I B ) and by the fact that during the preparation of the products of formula (I B ), the solid phase technique is combined to obtain the sequence 10-31 and the technique conventional in liquid phase for obtaining the sequence 1-9.
• the various amino acids are incorporated into the peptide chain using dicyclohexylcarbidimide (DCCI) and hydroxybenzotriazole (HOBT) by successively reacting protected amino acids in excess with respect to the proline first attached to the resin. The reaction is allowed to continue for two hours.
• the t-Bocs are eliminated by trifluoroacetic acid in solution in methylene chloride; after washing with methylene chloride, the neutralization is carried out with triethylamine or diisopropylethylamine (DIEA) in this same solvent.
• Asparagine and glutamine are introduced in the form of their paranitrophenyl esters, in solution in dimethylformamide (DMF).
• DMF dimethylformamide
• the control test for coupling with ninhydrin according to the method of Kaiser and Colescott is positive, treatment is carried out with pivalic acid (1%); if the blue color still persists in this test, the DMF / 1% AcOH is saturated with urea. However, even if this test is negative (no blue color), you can repeat each coupling twice systematically. If the coupling is not complete after 48 hours of reaction, as is the case for glutamine in position 20, the unreacted threonine is acetylated with acetylimidazole in chloride. methylene or by acetic acid and DCCI.
• the protective group of the imidazole is removed from the histidine, while the peptide is still bound to the resin, at pH 8, with mercapto-ethanol in solution in DMF.
• the peptide is separated from the resin by treatment with liquid hydrofluoric acid in the presence of anisole and methionine. All protecting groups are eliminated during this operation.
• the peptides are purified by gel filtration.
• the fractions corresponding to the main peaks are combined, lyophilized, taken up in a buffer at pH 8 and oxidized for 24 hours by a stream of air to form the disulfide bridge between the two cysteine residues.
• the fraction corresponding to the peak having the desired biological activity is purified again on ion exchange resin CMC 32 using an acid elution gradient.
• the peptide obtained is homogeneous on electrophoresis on paper and on chromatography on cellulose.
• sequence 10-31 of the products of formula (I B ) is carried out under the same conditions as those indicated above for the preparation of the products of formula (I A ). What has been indicated for glutamine in position 20 of the products of formula (I A ) is of course valid for position 19 of the products of formula (IB).
• the synthesis of the sequence 1-9 is carried out in a liquid medium.
• the Shard where A ' 2 has the above meaning and R ester, after cyclization, is condensed with the tripeptide A' 7 -Leu-Gly.
• Protecting groups generally used for the synthesis of peptides are used.
• the peptides are purified by gel filtration.
• the fractions corresponding to the main peaks are combined and lyophilized.
• the fraction corresponding to the peak having the desired biological activity is purified again on CMC 32 ion exchange resin by elution using a conductivity gradient.
• the peptide obtained after lyophilization appears homogeneous in electrophoresis on paper and in chromatography on cellulose.
• t-Boc amino acids carrying lateral functions originate from BACHEM (Dubendorf, Switzerland) or are prepared in the laboratory by the Schnabel technique (E. Schnabel, Liebigs Ann. Chem., 702, 188, 1967) in an autotitrator. Their purity is determined by polarimetry, melting point and chromatography on a thin layer of silica.
• the hydrolysis of the purified peptide is carried out in a vacuum-sealed tube in an HCI 6 N 110 medium. C, 24 hours).
• the apparatus used makes it possible to mechanically shake a 25 ml pyrex tube, the central compartment of which contains the resin (2 g). At the ends of the tube, 2 sintered glass filters are used for the introduction and then the elimination of the reagents (approximately 15 ml).
• the cycle 31 process is used by reacting 4 mmol of Boc-L-cysteine ether-S-p-methoxybenzyl (1.13 g).
• the peptide resin from cycle 3 previously washed with DMF (2x), is stirred (48 h) with 4 mmol of p-nitrophenyl ester of Boc-L-asparagine (1.45 g) in DMF medium, in the presence of HOBT (4 mmol).
• the resin is washed successively with DMF, CH 2 Cl 2 , CHsOH, CH 2 CI 2 .
• the reaction to ninhydrin is negative.
• hydrolysis of the purified peptide is carried out in a vacuum-sealed tube in HCl 6 N 110 medium. C, 24 h).
• the apparatus used makes it possible to mechanically shake a 25 ml pyrex tube, the central compartment of which contains resin (2 g). At the ends of the tube, 2 sintered glass filters are used for the introduction and then the elimination of the reagents (approximately 15 ml).
• A (5 g) is dissolved by cooling in TFA (15 ml). At room temperature, TFA is removed under vacuum and the residue dried under vacuum. It is dissolved in DMF (10 ml) and the pH brought to 6 using triethylamine (4 ml) while cooling. HOBT (5 g) and BOC-L-Ser (Bzl) OSu are then added and the mixture is stirred (3 days) at room temperature, at pH 6. N, N-dimethylamino-1,3-propanediamine is added, stirred (1 h), add H 2 0, extracted with AcOEt as described above. Cyclohexylamine (CHA) is added to dried AcOEt. Distillation under reduced pressure. The oily product B (4 g) is obtained.
• the activity is expressed in RCM units from the drop in calcemia, measured one hour after the intravenous injection of peptide to be tested suitably diluted in 0.1 N sodium acetate buffer pH 6 and 0.1% albumin , by comparison with the activity of "Research Standard B" suitably diluted.
• the biological activity of the peptide of Example 1 is greater than 4000 MRC units per mg and that of the peptide of Example 2 is greater than 4500 MRC units per mg.
• the usual dose which varies according to the product used, the subject treated and the condition in question, can for example be from 1 to 100 MRC units per day by the intramuscular or subcutaneous route.

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• Health & Medical Sciences (AREA)
• Chemical & Material Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Organic Chemistry (AREA)
• General Health & Medical Sciences (AREA)
• Medicinal Chemistry (AREA)
• Public Health (AREA)
• Toxicology (AREA)
• General Chemical & Material Sciences (AREA)
• Obesity (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Hematology (AREA)
• Pharmacology & Pharmacy (AREA)
• Diabetes (AREA)
• Animal Behavior & Ethology (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Engineering & Computer Science (AREA)
• Veterinary Medicine (AREA)
• Endocrinology (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Zoology (AREA)
• Gastroenterology & Hepatology (AREA)
• Biochemistry (AREA)
• Biophysics (AREA)
• Genetics & Genomics (AREA)
• Molecular Biology (AREA)
• Proteomics, Peptides & Aminoacids (AREA)
• Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
• Peptides Or Proteins (AREA)
• Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
• Compounds Of Unknown Constitution (AREA)
• Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
• Medicinal Preparation (AREA)

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Citations (2)

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• 1985
  • 1985-12-24 FR FR8519113A patent/FR2592049B1/fr not_active Expired
• 1986
  • 1986-12-19 WO PCT/FR1986/000440 patent/WO1987003884A1/fr active IP Right Grant
  • 1986-12-19 EP EP86402871A patent/EP0232650B1/fr not_active Expired
  • 1986-12-19 ES ES86402871T patent/ES2012456B3/es not_active Expired - Lifetime
  • 1986-12-19 AT AT86402871T patent/ATE48616T1/de not_active IP Right Cessation
  • 1986-12-19 DE DE8686402871T patent/DE3667472D1/de not_active Expired - Lifetime
  • 1986-12-19 JP JP62500164A patent/JPH0813838B2/ja not_active Expired - Lifetime
  • 1986-12-23 PT PT84011A patent/PT84011B/pt not_active IP Right Cessation
  • 1986-12-23 IE IE338686A patent/IE59489B1/en not_active IP Right Cessation
  • 1986-12-24 CA CA000526299A patent/CA1273750A/fr not_active Expired - Lifetime
• 1987
  • 1987-08-21 DK DK436187A patent/DK436187A/da not_active Application Discontinuation
  • 1987-08-21 FI FI873632A patent/FI87795C/fi not_active IP Right Cessation
• 1990
  • 1990-01-19 GR GR89400311T patent/GR3000279T3/el unknown

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